Using native fish in eco-genotoxic assessment of heavy metal contamination pollution arising from nearby large Brazilian rivers.

Worldwide water quality has declined progressively due to continuous pollution of aquatic resources by agrochemicals in particular heavy metals. Fish genotoxicity biomarkers are vital to identify and complement chemical parameters for determining environmental risk of adverse effects. Therefore, it was of interest to examine the eco-genotoxicity attributed to water pollution over different stream sections of Brazilian rivers by using Cichlasoma paranaense (Teleostei: Cichlidae), a neotropical freshwater cichlid fish, as a biological model. Chemical analysis of water and sediments collected from different Brazilian rivers sites demonstrated contamination by metals. Cichlasoma paranaense were collected at a reference location (a permanent water preservation area), maintained in the lab under standard conditions (controlled temperature, lighting, daily feeding, and constant aeration) and exposed to environmental samples of water and sediments. Subsequently, micronucleus (MN) and nuclear abnormalities (NA) frequencies were assessed in erythrocytes obtained from the caudal and gill regions. The highest concentrations of Cu were found in samples from river sites with forest fragmentation attributed to intensive agriculture practices. Similarly, exposure of fish to samples from agricultural areas induced significantly higher number of genotoxic effects. There was no marked difference between the tissues (tail and gill) regarding the observed frequencies of MN and NA. Thus C. paranaense fish served as a reliable model for detecting genotoxic effects, especially when water samples were collected near the discharge of agrochemicals. Evidence indicates that this method be considered for other global river sites which are also exposed to agrochemicals discharges containing Cu.

Assessment of the genotoxic potential of water courses impacted by wastewater treatment effluents using micronucleus assay in plants from the species Tradescantia.

Water pollution and the increase in genotoxic consequences in aquatic environments are well documented indicating the necessity and importance of biomonitoring programs. The objective of the present study was to determine the environmental quality of water resources and genotoxic potential of materials present within water samples obtained from the Perdizes River and the Mumbuca Stream, located in a region of discharge of wastewater treatment effluents using Tradescantia micronucleus assay (Trad - MCN). Water samples were collected from different locations up and downstream of the wastewater treatment plant during rainy season and subsequently submitted to physico-chemical analysis and Trad-MCN bioassay. The spatial distribution of the physico-chemical parameters assessed suggested that discharges of wastewater treatment effluents reduced water quality at all sites examined. Further, exposure to wastewater treatment effluents produced genotoxic effects on tetrads of Tradescantia pallida. These results reinforce the sensitivity of the Trad-MCN bioassay and its potential application in water quality monitoring programs concomitant with physicochemical evaluation.

In situ assessment of the paraguay river water, in Brazilian Pantanal, by means of micronucleus assay with fish and chemical analysis.

The aim of this research was to assess water quality in a stretch of the Paraguay River within the Brazilian Pantanal by means of a micronucleus assay with fish, and by water and sediment physicochemical analysis. Significant increases (p >0.05) in the frequency of micronuclei (MN) and micronucleated cells (MNC) occurred in erythrocytes of Pimelodus maculatus and Leporinus friderici at two river sites in the town of Caceres relative to an upstream reference site. The results demonstrate that the Paraguay River water near Caceres has been receiving genotoxic effluents, which may be associated with the presence of chromium, sulfides, oil and grease, and/or other chemicals.

Photochemical Properties of trans-[Ru(NH3 )4 (bpa)(L)]2+ (L = py, isn, 4-acpy or 4-pic).

Photochemical reactions of ruthenium (II) complexes of type trans-[Ru(NH3 )4 LL']2+ , where L is a nitrogenous heterocyclic ligand, pyridine (py), isonicotinamide (isn), 4-acetylpyridine (4-acpy) or 4-picoline (4-pic), and L´ is a 1,2-bis(4-pyridyl)ethane (bpa) ligand, were studied with the purpose of evaluating the ligand exchange when, in solution, the complexes are irradiated at the wavelengths of 365, 436, 480 and 519 nm. The study revealed that at lower wavelengths, a labilization process is observed for py and 4-pic ligands, even at low quantum yields, indicating the dependence of the photolabeling process on the wavelength. The study also reveals that for the filters of greater wavelength, the processes of photolabilization do not occur for any of the studied complexes. The study also shows that there are no photolization processes for the complexes obtained with the isn and 4-acpy ligands, and it is therefore possible to classify them as nonreactive.

Evaluation of toxicity, mutagenicity and carcinogenicity of samples from domestic and industrial sewage.

Physico-chemical and toxicological analyses are of fundamental importance to determine water quality. The objectives of the present study were to evaluate the toxicity, mutagenicity and carcinogenicity of samples from the Mumbuca Stream and the Perdizes River, through both SMART and the wts test, respectively, in somatic cells of Drosophila melanogaster and to quantify the amount of heavy metals and other pollutants, which are indicative of environmental quality. Water samples were collected (M1, M2, P1, P2 and MP) and submitted to physico-chemical analysis, calculating the water quality index for each sampling site. In order to evaluate the toxicity, mutagenicity and carcinogenicity of the samples, third instar larvae descended from the crossing between virgin female wts/TM3, sb1 and mwh/mwh males (wts test) and ST and HB (SMART) crosses were treated with samples from P1, P2, M1, M2 and MP sites. The physico-chemical analysis and the biological assay allowed us to conclude that undetected values for heavy metals and the low frequency of mutant spots (SMART) and epithelial tumor (wts) in treated flies from the Mumbuca Stream and Perdizes River may be due to the reduction of ceramic activities in the municipality. The physico-chemical analyzes identified altered the environmental quality parameters, which directly influenced the survival of D. melanogaster treated with samples of M2 and MP, which according to the WQI were classified as regular and poor environmental quality, respectively. The altered parameters may be due to clandestine domestic sewage sent downstream of the effluent.

Assessing the impact of pollution on the Japaratuba river in Brazil using the Drosophila wing spot test.

The Drosophila melanogaster somatic mutation and recombination test (SMART) was used to assess the genotoxicity of surface (S) and bottom (B) water and sediment samples collected from Sites 1 and 2 on the Japaratuba River (Sergipe, Brazil), an area impacted by a petrochemical industrial complex that indirectly discharges treated effluent (produced water) into the river. The genotoxicity tests were performed in standard (ST) cross and high bioactivation (HB) cross flies and were conducted on samples taken in March (dry season) and in July (rainy season) of 2003. Mutant spot frequencies found in treatments with unprocessed water and sediment samples from the test sites were compared with the frequencies observed for similar samples taken from a clean reference site (the Jacarecica River in Sergipe, Brazil) and those of negative (ultrapure water) controls. While samples from the Japaratuba River generally produced greater responses than those from the Jacarecica River, positive responses were detected for both the test and reference site samples. All the water samples collected in March 2003 were genotoxic. In July 2003, the positive responses were restricted to water samples collected from Sites 1 B and 2 S in the ST cross. The genotoxicity of the water samples was due to mitotic recombination, and the samples produced similar genotoxic responses in ST and HB flies. The spot frequencies found in the July water samples were considerably lower than those for the March water samples, suggesting a seasonal effect. The only sediment samples that were genotoxic were from Site 1 (March and July) and from the Jacarecica River (March). The genotoxins in these samples produced both somatic mutation (limited to the Site 1 sample in HB flies) and recombination. The results of this study indicate that samples from both the Japaratuba and Jacarecica Rivers were genotoxic, with the most consistently positive responses detected with Site 1 samples, the site closest to the putative pollution source.

Biological monitoring and B chromosome frequency in Bagre (Rhamdia quelen) in southeast Brazil.

The genus Rhamdia presents B chromosomes which appear to be present in most species of the genus and thus represent an important characteristic in the evolutionary process. Furthermore, variations in environmental conditions can induce the presence of B chromosomes generated by alterations in the cell cycle, due to the interference from pollutants. The present study aimed to evaluate the cytogenetic aspects of individuals of a population of Rhamdia quelen collected in three areas with differing standards of water quality in the River Uberabinha, a region of the County of Uberlandia, Minas Gerais, Brazil. The Piscine Micronucleus Test results indicate significant genotoxic and cytotoxic potential at the sampling Sites. The chromosome count yielded the modal number 2n=58 with variance between zero and seven B chromosomes. The highest frequency of B chromosomes and the presence of karyotypes with seven supernumerary chromosomes occurred at Site 3, referring, thus, to the location of the highest genotoxic potential. There was a positive correlation between the presence of B chromosomes and the reduction in environmental quality. Therefore, the process of bioaccumulation of heavy metals in aquatic environments may be crucial to determine the presence of B chromosomes.

Cytotoxic effects of the compound cis-tetraammine(oxalato)ruthenium(III) dithionate on K-562 human chronic myelogenous leukemia cells.

Chemotherapy is a common treatment for leukemia. Ruthenium complexes have shown potential utility in chemotherapy and photodynamic therapy. The identification of new chemotherapeutics agents is critical for further progress in the treatment of leukemia. Ruthenium complexes generally have lower toxicities compared to cisplatin attributed to their specific accumulation in cancer tissues. Based on these evidences, in the present work we studied the cytotoxic activity of the ruthenium(III) compound cis-tetraammine(oxalato)ruthenium(III) dithionate - {cis-[Ru(C2O4)(NH3)4]2(S2O6)} against human chronic myelogenous leukemia cells (K-562) tumor cell line. The tested compound induces cell death in a dose and time dependent manner on K-562 cells. It is found that the effect was improved linearly while prolonging the incubation time. Compared to the cell cycle profiles of untreated cells, flow cytometric analysis indicated the sub-G1 arresting effect of ruthenium compound on K-562 cells. In our study, {cis-[Ru(C2O4)(NH3)4]2(S2O6)} shows a significant increase in tailed cells in any of the concentrations tested compared with negative control. Consequently, the concentration of {cis-[Ru(C2O4)(NH3)4]2(S2O6)} might be associated cytotoxicity with direct effect on K-562 cells DNA. Thus, it can be deducted that ruthenium-based compounds present selectivity to enter both tumor and normal cells. Additional studies are needed to determine the molecular mechanisms of the active components and to evaluate the potential in vivo anticancer activity of the cis-tetraammine(oxalato)ruthenium(III) dithionate.

Induction of cell cycle arrest and apoptosis by ruthenium complex cis-(dichloro)tetramineruthenium(III) chloride in human lung carcinoma cells A549.

Lung cancer is one of the leading causes of death in the world, and non-small cell lung carcinoma (NSCLC) accounts for approximately 75-85% of all lung cancers. In the present work, we studied the cytotoxic activity, cell cycle arrest and induction apoptosis of the compound cis-(dichloro)tetramineruthenium(III) chloride {cis-[RuCl(2)(NH(3))(4)]Cl} in human lung carcinoma tumor cell line A549. The results of MTT and trypan blue assays showed that cis-[RuCl(2)(NH(3))(4)]Cl causes reduction in the viability of A549 cells when treating with 95 and 383 µM of the compound for 48 and 72 h. Lower concentrations of the compound (19, 3.8 and 0.38 µM), however, only slightly affected cell viability. The IC(50) value for the compound was about 383 µM. Survival analysis of the A549 cells after treatment with ruthenium(III) compound using long term clonogenic assay showed that it reduced colony formation ability at concentrations of 0.38 and 3.8 µM, and at concentrations of 95 and 383 µM no colonies were observed. Cell cycle analysis showed that compound ruthenium led to an accumulation of A549 cells in S phase and increased in the sub-G1 peak. In addition, cis-(dichloro)tetramineruthenium(III) chloride treatment induced apoptosis, as observed by the increased numbers of annexin V-positive cells and increased messenger RNA expression of caspase-3.

The ruthenium complex cis-(dichloro)tetrammineruthenium(III) chloride induces apoptosis and damages DNA in murine sarcoma 180 cells.

Ruthenium (III) complexes are increasingly attracting the interest of researchers due to their promising pharmacological properties. Recently, we reported that the cis-(dichloro)tetrammineruthenium (III) chloride compound has cytotoxic effects on murine sarcoma 180 (S-180) cells. In an effort to understand the mechanism responsible for their cytotoxicity, study we investigated the genotoxicity, cell cycle distribution and induction of apoptosis caused by cis- (dichloro) tetrammineruthenium (III) chloride in S-180 tumour cells. cis-(dichloro) tetrammineruthenium (III) chloride treatment induced significant DNA damage in S-180 cells, as detected by the alkaline comet assay. In the cell cycle analysis, cis-(dichloro) tetrammineruthenium (III) chloride caused an increase in the number of cells in G1 phase, accompanied by a decrease in the S and G2 phases after 24 h of treatment. In contrast, the cell cycle distribution of S-180 cells treated with cis-(dichloro) tetrammineruthenium (III) chloride for 48 h showed a concentration-dependent increase in the sub-G1 phase (indicating apoptosis), with a corresponding decrease in cells in the G1, S and G2 phases. In addition, cis-(dichloro) tetrammineruthenium(III) chloride treatment induced apoptosis in a time-dependent manner,as observed by the increased numbers of annexin V-positive cells. Taken together, these findings strongly demonstrate that DNA damage, cell cycle changes and apoptosis may correlate with the cytotoxic effects of cis-(dichloro) tetrammineruthenium (III) chloride on S-180 cells.

The ruthenium complex cis-(dichloro)tetraammineruthenium(III) chloride presents selective cytotoxicity against murine B cell lymphoma (A-20), murine ascitic sarcoma 180 (S-180), human breast adenocarcinoma (SK-BR-3), and human T cell leukemia (Jurkat) tumor cell lines.

The aim of present study was to verify the in vitro antitumor activity of a ruthenium complex, cis-(dichloro)tetraammineruthenium(III) chloride (cis-[RuCl(2)(NH(3))(4)]Cl) toward different tumor cell lines. The antitumor studies showed that ruthenium(III) complex presents a relevant cytotoxic activity against murine B cell lymphoma (A-20), murine ascitic sarcoma 180 (S-180), human breast adenocarcinoma (SK-BR-3), and human T cell leukemia (Jurkat) cell lines and a very low cytotoxicity toward human peripheral blood mononuclear cells. The ruthenium(III) complex decreased the fraction of tumor cells in G0/G1 and/or G2-M phases, indicating that this compound may act on resting/early entering G0/G1 cells and/or precycling G2-M cells. The cytotoxic activity of a high concentration (2 mg mL(-1)) of cis-[RuCl(2)(NH(3))(4)]Cl toward Jurkat cells correlated with an increased number of annexin V-positive cells and also the presence of DNA fragmentation, suggesting that this compound induces apoptosis in tumor cells. The development of new antineoplastic medications demands adequate knowledge in order to avoid inefficient or toxic treatments. Thus, a mechanistic understanding of how metal complexes achieve their activities is crucial to their clinical success and to the rational design of new compounds with improved potency.

The ruthenium complex cis-(Dichloro)Tetraammineruthenium(III) chloride presents immune stimulatory activity on human peripheral blood mononuclear cells.

Ruthenium compounds in general are well suited for medicinal applications. They have been investigated as immunosuppressants, nitric oxide scavengers, antimicrobial agents, and antimalarials. The aim of this study is to evaluate the immunomodulatory activity of cis-(dichloro)tetraammineruthenium(III) chloride (cis-[RuCl(2)(NH(3))(4)]Cl) on human peripheral blood mononuclear cells (PBMC). The cytotoxic studies performed here revealed that the ruthenium(III) complex presents a cytotoxic activity towards normal human PBMC, only at very high concentration. Results also showed that cis-[RuCl(2)(NH(3))(4)]Cl presents a dual role on PBMC stimulating proliferation and interleukin-2 (IL-2) production at low concentration and inducing cytotoxicity, inability to proliferate, and inhibiting IL-2 production at high concentration. The noncytotoxic activity of cis-[RuCl(2)(NH(3))(4)]Cl at low concentration towards PBMC, which correlates with the small number of annexin V positive cells and also the absence of DNA fragmentation, suggest that this compound does not induce apoptosis on PBMC. For the first time, we show that, at low concentration (10-100 microg L(-1)), the cis-[RuCl(2)(NH(3))(4)]Cl compound induces peripheral blood lymphocytes proliferation and also stimulates them to IL-2 production. These results open a new potential applicability of ruthenium(III) complexes as a possible immune regulatory compound acting as immune suppressor at high concentration and as immune stimulator at low concentration.

The compound cis-(dichloro)tetrammineruthenium(III) chloride induces caspase-mediated apoptosis in K562 cells.

Ruthenium(III) complexes are increasingly attracting the interest of researchers due to their promising pharmacological properties. In the present study, we investigated the ability of cis-(dichloro)tetrammineruthenium(III) chloride to produce lethal effects in human chronic myelogenous leukemia K562 cells. The MTT tetrazolium reduction test and the trypan blue exclusion assay revealed that the IC(50) for the compound after 48 h of incubation with K562 cells was approximately 10.74 and 73.45 microM, respectively. Interestingly, it was observed that this compound exhibits mild cytotoxicity towards MRC-5 human fibroblast cells (IC(50)>383 microM). Flow cytometric analysis revealed that cis-(dichloro)tetrammineruthenium(III) chloride was capable of change cell cycle distribution since the percentage of cells in the G1, S and G2 phases decreased. In addition, treatment with this compound induced apoptotic cell death in K562 cells, demonstrated by increased DNA content in the sub-G1-peak and a significant increase in caspase-3 activity. Assay using cyclosporin A, an inhibitor of the mitochondrial permeability transition pore (MPT) showed that the preincubation of K562 cells with this inhibitor had not effect on cis-(dichloro)tetrammineruthenium(III) chloride induced caspase-3 activation. In summary, cis-(dichloro)tetrammineruthenium(III) chloride displayed a significant cytotoxic effect through cell cycle arrest and apoptotic induction in K562 cells, which suggests that cis-(dichloro)tetrammineruthenium(III) chloride might have therapeutic potential against leukemia.

Cytotoxic and genotoxic effects of cis-tetraammine(oxalato)ruthenium(III) dithionate on the root meristem cells of Allium cepa.

Ruthenium complexes have attracted much attention as possible building blocks for new transition-metal-based antitumor agents. The present study examines the mitotoxic and clastogenic effects induced in the root tips of Allium cepa by cis-tetraammine(oxalato)ruthenium(III) dithionate {cis-[Ru(C(2)O(2))(NH(3))(4)](2)(S(2)O(6))} at different exposure durations and concentrations. Correlation tests were performed to determine the effects of the time of exposure and concentration of ruthenium complex on mitotic index (MI) and mitotic aberration index. A comparison of MI results of cis-[Ru(C(2)O(2))(NH(3))(4)](2)(S(2)O(6)) to those of lead nitrate reveals that the ruthenium complex demonstrates an average mitotic inhibition eightfold higher than lead, with the frequency of cellular abnormalities almost fourfold lower and mitotic aberration threefold lower. A. cepa root cells exposed to a range of ruthenium complex concentrations did not display significant clastogenic effects. Cis-tetraammine(oxalato)ruthenium(III) dithionate therefore exhibits a remarkable capacity to inhibit mitosis, perhaps by inhibiting DNA synthesis or blocking the cell cycle in the G2 phase. Further investigation of the mechanisms of action of this ruthenium complex will be important to define its clinical potential and to contribute to a novel and rational approach to developing a new metal-based drug with antitumor properties complementary to those exhibited by the drugs already in clinical use.

Mutagenic and genotoxic effects of cis-(dichloro)tetraammineruthenium(III) chloride on human peripheral blood lymphocytes.

Chemotherapeutic agents play an important role in cancer treatment mostly due their systemic action on human organism allowing access to liquid tumors and even metastases. Among these drugs, ruthenium compounds have been showing promising results to treat tumors and represent an important development of new antitumor therapy. This study presents the evaluation of cis-(dichloro)tetraammineruthenium(III) chloride, cis-[RuCl(2)(NH(3))(4)]Cl, genotoxic effects using human peripheral blood lymphocytes cultured in vitro. Mitotic index (MI), chromosome aberrations (CA), and DNA damage using the comet assay were analyzed. MI in human peripheral blood lymphocyte cultures treated with 1, 10, 100, and 1,000 microg mL(-1) cis-[RuCl(2)(NH(3))(4)]Cl were 5.9%, 4.6%, 3.9%, and 0%, respectively. Doxorubicin chloridate was used as the positive control. CA derived from 1, 10, and 100 microg mL(-1) concentrations were defined as spontaneous when compared with the negative control, and at the concentration of 1,000 microg mL(-1), the cell cycle was inhibited (IM = 0%). Results obtained for the comet assay using cis-[RuCl(2)(NH(3))(4)]Cl suggest that this compound has no genotoxic activity against cultured human peripheral blood lymphocytes.

Genotoxicity of water from the Paraguay River near Cáceres-MT, Brazil in the Drosophila wing-spot test.

The genotoxic activity of surface water samples from four sites along the Paraguay River, near Cáceres, Mato Grosso State, Brazil, was investigated using the Drosophila melanogaster Somatic Mutation and Recombination test (SMART). Effluents from sanitary sewers and agroindustrial effluents (residual effluents from slaughterhouses, leather tanneries, and dairies) flow into the Paraguay River, and directly or indirectly contaminate water from sampling sites 1-3. Site 4 was an upriver reference site that received no domestic or agroindustrial discharges. Water was collected at 4 time periods: September 2003 and August 2004 (periods of low water or drought); and April 2004 and March 2005 (periods of high water or flood). Chromium concentrations above statutory limits were detected at sites 1-3 (August 2004), and sites 1, 2 and 4 (March 2005). Sulfur compounds were also detected at sites 1-3. The SMART performed using standard (ST) cross flies detected genotoxic responses in only two samples, the August 2004 site 1 sample and the March 2005 site 2 sample. Many more samples were positive using high bioactivation (HB) cross flies: site 1 (all collection periods), site 2 (September 2003 and April 2004), and site 3 (September 2003 and August 2004). Mutant frequency comparisons between marker-heterozygous and balancer-heterozygous flies from the HB cross indicated that the positive genotoxic responses for the site 2 (April 2004) and site 3 (September 2003) samples were due mainly to mitotic recombination. Our findings indicate that the section of the Paraguay River within the urban perimeter of Cáceres is contaminated with genotoxic agents.

Determination of chromium (III) using a homogenous mixture of water-ethanol-methylisobutylketone solvents.

The use of organic solvents to increase metal ion determination sensitivity by atomic absorption spectrophotometry with flame is quite common. The most employed organic solvent is 4-methyl-2-pentanona (methylisobutylketone, MIBK) which optimizes sample vaporization and combustion. In this work, we present the use of a homogeneous mixture of water-ethanol-MIBK solvents (1:14:10 v/v, respectively), named the single-phase solution instead of employing pure organic solvents to determine chromium (III) ions by atomic absorption spectrophotometry with flame. The analytical calibration curve in single-phase solution evaluated up to 8 mugml(-1) was linear and was described as Abs=0.0048 C(Cr(III))-0.0010 (r(2)=0.9998). Stability in the measurement as well as an increase in sensitivity more than twice as high when compared to determinations exclusively made in aqueous solutions were observed. The exactness of the determinations was evaluated with the same steel standards.

Análise espectroscópica do conteúdo de monômero residual liberado pelas resinas acrílicas termopolimerizáveis / Spectroscopic analysis of the content of residual monomer released by the heat polymerized acrylic resins

As técnicas espectroscópicas ultravioleta (UV) e infravermelho (IR) foram utilizadas neste trabalho com o objetivo de analisar o conteúdo de monômero residual em duas resinas acrílicas termopolomerizáveis de uso odontológico, com relaçäo a variaçäo nos ciclos de polimerizaçäo e nos tempos de dosagem do monômero. Os resultados obtidos mostraram que as quantidade de monômero residual liberadas pelas resinas estudadas foram baixas e dependentes do tempo e temperatura de polimerizaçäo. A liberaçäo do monômero também foi maior nas primeiras horas, diminuindo a medida que decorria o tempo de imersão em água